Methods of Treating A Bacterial Infection using F. Tularensis Lipopolysaccharide
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[0061] Subjects and Reagents
[0062] Five to 6-wk-old C57BL / 6J mice were purchased from the Jackson Laboratory (Bar Harbor, Me.). Mice were injected i.p. with purified Ft LVS LPS or inoculated i.p. or i.d. with Ft LVS bacteria. At the indicated times after inoculation, mice were sacrificed and livers, lungs, spleen, and blood were collected. Organs were snap frozen in an ethanol / dry ice bath and then stored at −80° C. for subsequent RNA extraction. Serum samples were collected and stored at −80° C. Serum cytokine concentrations were determined using ELISA assays or the Luminex® 100™ Total System (Luminex Corporation, Austin, Tex.) by the Cytokine Core Facility (Univ. of Maryland, Baltimore).
[0063] Peritoneal macrophages were isolated from mice 4 days after i.p. injection of sterile 3% thioglycollate as described previously in Salkowski, C. A., et al., J. Immunol. 163:1529-1536 (1999). Briefly, cells were washed in sterile 1×PBS and then resuspended in RPMI 1640 (Invitrogen, Carlsbad...
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